Determination of erythrocyte porphyrins by EPI-illumination fluorescence microscope with capillary electrophoresis

Abstract

A micellar electrokinetic capillary chromatographic (MEKC) method is described for the simultaneous determination of zinc protoporphyrin (ZnPP) and protoporphyrin (PP) in whole blood. Reproducibility of migration times and peak areas of ZnPP and PP have been shown to improve significantly with the use of an internal standard. Erythrocyte porphyrins were extracted from whole blood with N,N-dimethylformamide (DMF), followed by sample cleanup and preconcentration on C-18 cartridges. Analytical recoveries of the porphyrins were better than 88%. An epi-illumination fluorescence microscope was used for the detection of these porphyrins. Limits of detection were found to be 50 nM and 8 nM for ZnPP and PP, respectively.